|Publication number||US4517288 A|
|Application number||US 06/227,664|
|Publication date||May 14, 1985|
|Filing date||Jan 23, 1981|
|Priority date||Jan 23, 1981|
|Also published as||CA1179940A, CA1179940A1, DE3277704D1, EP0070300A1, EP0070300A4, EP0070300B1, WO1982002601A1|
|Publication number||06227664, 227664, US 4517288 A, US 4517288A, US-A-4517288, US4517288 A, US4517288A|
|Inventors||Joseph L. Giegel, Mary M. Brotherton|
|Original Assignee||American Hospital Supply Corp.|
|Export Citation||BiBTeX, EndNote, RefMan|
|Patent Citations (18), Non-Patent Citations (2), Referenced by (283), Classifications (14), Legal Events (16)|
|External Links: USPTO, USPTO Assignment, Espacenet|
1. Field of Invention
The present invention relates to a solid phase system for ligand assay. More particularly, the invention relates to an immunological method for analyzing biological fluids wherein analytical reactions are conducted in an inert porous medium.
2. Discussion of Prior Art
Variations of natural immunological reactions have been found very useful as analytical techniques. These reactions have been most useful in clinical laboratory procedures, but their use is not limited to clinical applications. Because of the specificity of the reactions, they are of particular advantage in analyzing complex biological fluids. Often, conventional chemical analyses are not capable of differentiating complex molecules in a biological fluid. Such fluids can be analyzed for a variety of components such as drugs, enzymes, hormones, etc. by contacting the fluid with an appropriate antibody. Likewise, analyses for specific antibodies can be conducted by contacting the fluid with an appropriate antigen. Other natural binding proteins are also quite useful in some types of assay procedures.
Unfortunately, the antibody-antigen reaction is generally not directly measurable, therefore, techniques have been devised for its indirect measurement. For instance, an antibody, antigen, or binding protein (collectively referred to as a ligand) may be labeled by various means. The amount of bound labeled ligand can thus be correlated to the concentration of the analyte in the biological fluid. Conventional labels include radioactive tags, e.g., 125 I or tritium, enzymes, chromophores, fluorophores and enzymes cofactors and effectors. In the case of radioactive tags, the concentration of the labeled ligand is usually determined by placing the compound in a scintillation counter. Enzymes may be measured by reacting the labeled ligand with a substrate, which by the action of the enzyme, releases a chromogenic or fluorogenic substance that can be measured by conventional techniques. Ligands labeled with enzyme cofactors or effectors can be detected similarly by their effect on enzyme action on a substrate. Compounds labeled with chromophores may be directly measurable, e.g., by fluorescence, ultraviolet spectroscopy or other spectroscopic means.
Immunochemical assays generally fall into one of two classifications. In the competitive assay, a limited quantity of binding material is contacted with a solution containing the analyte and a known concentration of a labeled analyte. The labeled and unlabeled analyte compounds compete for the binding sites on the binding material. By reference to a calibration curve, the amount of labeled analyte bound to the binding material can be correlated with the concentration of the analyte in the test solution. A second type of immunological assay, the sandwich assay, involves contacting a binding material with a solution containing the analyte to cause the analyte to bind to the binding material. This complex is then contacted with a solution of a labeled binding material, generally an antibody, which reacts with the bound analyte. The amount of bound labeled binding material is thus directly proportional to the amount of bound analyte.
In all of the described methods, an essential step is to separate the unbound labeled material from the bound labeled material. A technique widely employed for such separation is to immobilize one of the reactants. For instance, an antibody may be adsorbed onto a solid support such as a test tube wall. After labeled material and analyte become bound to the immobilized antibody, the solid support is rinsed free of unbound labeled material. A variety of solid supports have been proposed for this purpose. Such supports include test tube walls, plastic cups, beads, plastic balls and cylinders, paper, and glass fibers.
In U.S. Pat. No. 3,888,629, June 10, 1975, K. D. Bagshawe discloses a reaction cell and an immunoanalytical method, in which antibody is immobilized in a matrix pad of absorbent material such as a glass fiber pad. The reference discloses an antibody impregnated sheet of the absorbent material. Disks or pads of the material are then punched from the mat and placed in a reaction cell. Solutions of the analyte and other reactants are placed on the pad where the immunological reaction occurs. A buffer is then filtered through the pad to wash out unreacted labeled material. Because of surface tension and capillary action, liquid does not easily pass through the pad; therefore, an absorbent material is placed under the pad to facilitate filtration.
To quantitate the reaction conducted in the Bagshawe cell, the pad must be removed from the cell and either placed in a gamma counter (in the case of a radioimmunoassay) or placed in some type of indicator solution (in the case of an enzymeimmunoassay).
A need exists for a rapid, quantitative solid phase ligand assay, which can be conducted entirely on a solid matrix. Advantageously, the method would not require special reaction cells, and the separation of unreacted labeled material could be effected cleanly without filtering large amounts of solvents through the matrix. The amount of binding material deposited in the reaction zone on the solid matrix should be accurately controllable. Such accurate control has not heretofore been realized in prior art methods.
In accordance with the present invention, a method is disclosed for conducting a ligand assay in an inert porous medium wherein a binding material is immobilized within said medium, the improvement which comprises:
(a) immobilizing a binding material within a finite zone of said medium;
(b) applying to said zone under binding conditions an analyte to which said binding material is specific;
(c) applying to said zone a labeled indicator under conditions which allow said labeled indicator to become immobilized within said zone in an amount which can be correlated to the amount of analyte in said zone;
(d) applying to substantially the center of said zone a stream of solvent in a quantity sufficient to effect a radial chromatographic separation of unbound labeled indicator from said zone; and
(e) determining the amount of labeled indicator remaining in said zone.
The method of the present invention is conducted in a porous solid matrix. Such matrix is in sheet form and may be made of any suitable inert material which does not deleteriously react with any of the reactants, the products, or the solvents. The term, inert, when referring to the matrix, means that the matrix does not chemically react with any of the materials applied thereto, does not dissolve, does not react immunologically with such materials, and has negligible nonspecific attraction for such materials. Thus, the matrix, or more precisely, the interstices within the matrix, simply acts as a vessel or site for reactions to occur.
The interstices or pores within the matrix are small enough so that the reaction fluid is retained within the matrix by capillary action. The matrix is advantageously a mat of compressed fibers, such as a mat of glass or synthetic fibers or a porous paper mat. The matrix may, however, be constructed of other porous materials such as sintered glass, ceramics, synthetic spongy materials, etc. A glass fiber mat is the preferred matrix because of its inertness. Other materials such as paper tend to have a greater non-specific attraction for the materials applied thereto and may require pretreatment to inactivate reactive sites.
The present invention is useful for a wide variety of biological assays. For instance, blood or urine may be quickly and accurately analyzed for therapeutic drugs, natural or synthetic steroids, hormones, antibodies and the like. Therapeutic drugs which may be determined by this method include digoxin, dilatin, phenobarbital, theophylline, gentamycin, guinidine, propranolol, and the like. Steroids, such as cortisol, aldosterone, testosterone, progesterone and estriol may also be monitored conveniently by the present method. The method may be useful for determining serum or urine levels of hormones such as the thyroid hormones, triiodo-thyronine; the peptide hormones, insulin, coricotropin, gastrin, angiotensin, and proangiotensin; the polypeptide hormones, thyrotropin, luteotropin, and somatotropin, and the like. The binding materials for such assays are generally antibodies specific for the analyte of interest. Such antibodies may be prepared by conventional methods, well known by those skilled in the art. Typically, antibodies are prepared by innoculating an animal, such as rabbit, goat, horse, donkey, sheep, chicken, or human with the antigen of interest, and harvesting the antibodies from the blood of the animal. In addition to antibodies, the binding material may be a non-antibody binding protein, such as vitamin B-12 intrinsic factor, thyroxine binding globulin, folate binding protein, or sex hormone binding protein.
A first step of the present method is to immobilize a binding material within a finite zone of the matrix. Immobilization can be accomplished by any convenient method, such as adsorption, evaporative deposition from a volatile solvent solution, covalent bonding between the binding material and the matrix, or immunological immobilization. Covalent bonding may, for example, involve bonding the binding material to a matrix through a coupling agent, such as cyanogen bromide or glutaraldehyde, as described by Grubb, et al., U.S. Pat. No. 4,186,146 (Sept. 18, 1979). Immunological immobilization is preferred for the practice of the present invention. To immunologically immobilize the binding material within the matrix, the binding material can be reacted with an antibody to such material in a solution absorbed in the reaction zone. In practice, the immunological immobilization may be accomplished by applying to a point on the matrix a small volume, e.g., 2-100 μl, of a mixture of a solution of the binding material and an antiserum or a solution thereof containing an antibody to the binding material. By capillary action, the mixture diffuses out from the point of application to encompass a finite zone of the matrix. The matrix is then incubated under conditions of temperature and humidity for a period of time sufficient to effect an immunological reaction between the reactants. The reaction conditions will vary depending upon the reactants being used, but generally a temperature of from about -10° C. to about 60° C., preferably from about 0° to about 25° C. is employed, and an incubation time of from about 1 minute to about 24 hours, preferably from about 3 minutes to about 18 hours has been found sufficient to effect the reaction.
For quantitative applications of the present method, the amount of binding material immobilized within the reaction zone should be accurately controlled. One of the advantages of the present method over those described in the prior art is that very accurate control can be achieved through the use of accurate volumetric micropipettes and solutions of known concentrations. The antiserum is generally employed in an amount to insure that substantially all of the binding material is immobilized.
Alternative methods of immobilizing the binding material in the matrix include applying solutions of binding material and antibody to the reaction zone, either sequentially or simultaneously, and incubating to effect the immunological reaction. The matrix sheet may also be impregnated with the antibody by applying a solution thereof uniformly to the sheet and then drying the sheet. As the solution of binding material is spotted on the matrix, the antibody in the reaction zone is redissolved and reacts immunologically with the binding material.
Those skilled in the art will appreciate that a sheet of matrix may have one reaction zone or several. The sheet may be dried after immobilization of the binding material, and stored for prolonged periods before being utilized in an assay procedure. Sheets having multiple reaction zones may have several spots of the same binding material or may comprise different binding materials, for different assays.
Solutions of analyte and labeled indicator are applied to the reaction zone containing the immobilized binding material; and after incubation under appropriate conditions, the immunological reaction occurs resulting in analyte and labeled indicator being bound in the reaction zone. Whether the analyte is applied prior to, concurrently with, or subsequent to the application of the labeled indicator depends upon the type of assay being conducted. For instance, in a competitive assay for an antigen, such as the drug digoxin, the binding material is an anti-digoxin antibody. The analyte, which may be a patient serum, and the labeled indicator, which is suitably labeled digoxin, are applied to the reaction zone substantially concurrently. In a sandwich assay, e.g., for IgG, the labeled indicator would be a labeled anti-IgG antibody, and could be applied to the reaction zone before, after or simultaneously with the reaction between the analyte and the binding material.
The concentrations of analyte and labeled indicator are controlled so that the amounts of such compounds deposited in the reaction zone are appropriate for the particular type of reaction being employed. Such concentrations can be determined in accordance with relationships known in the art. For instance, in a competitive assay, the total amount of analyte and labeled indicator applied to the reaction zone is substantially equal to or greater than that required to bind to all of the binding sites on the binding material. The total concentration of binding sites of the analyte and labeled indicator generally constitute from about 0.1 to about 10, preferably from about 0.5 to about 1.5 times the concentration of binding sites on the binding material. In a sandwich assay, the concentration of the analyte will be such that it can be substantially completely bound in the reaction zone. To accomplish substantially complete binding, generally there is an excess of binding material in the reaction zone with respect to the analyte. The binding material is advantageously employed at a concentration of from about 1 to about 100, preferably about 1 to 10, times that required to bind all of the binding sites of the analyte. The labeled indicator should then be employed in a quantity sufficient to react with the bound analyte. Thus, the labeled indicator is employed in an amount from about 1 to 2, preferably 1 to 1.5, times the amount required to react with all of the bound analyte.
The analyte and labeled indicator are applied to the reaction zone in solution in such a manner that they diffuse throughout substantially the entire reaction zone. Such application is advantageously accomplished by pipetting small volumes of solutions to substantially the center of the zone.
If desired, matrix sheets can be prepared for routine use which have binding material and labeled indicator already applied thereto. The binding material may be applied as described above. The labeled indicator may then be applied to the surface of the matrix, e.g., by a printing process. Advantageously, a barrier layer is applied to the surface prior to application of the labeled indicator to prevent premature reaction of the labeled indicator with the binding material. The barrier layer is a thin coating of a material which is water soluble. Thus, when the analyte is applied to the reaction zone, the barrier layer dissolves and allows the labeled indicator to diffuse into the reaction zone.
The matrix sheet is incubated under conditions which enable the analyte and/or the labeled indicator to be bound to the binding material. A principal advantage of the present invention is that such reactions typically occur very rapidly at room temperature. Generally, incubation temperatures of from about 5° C. to about 60° C. are employed with a preferred temperature in the range of from about 15° to about 40° C. The immunological reactions are usually sufficiently complete in from about a few seconds to about 30 hours, depending on the particular reaction being conducted.
After the reactions have occurred, the unbound labeled indicator is separated from the reaction zone. In accordance with the present invention, the unbound labeled indicator is quickly and quantitatively separated from the reaction zone by a chromatographic procedure. A stream of a solvent, in which the labeled indicator is soluble, is applied to substantially the center of the reaction zone. The solvent may be water or a buffer solution in which such compounds are conveniently dissolved. As the solvent migrates radially out from the center of the reaction zone, unbound reactants are chromatographically separated from the bound reactants. A small quantity of such solvent effectively separates the unbound reactants from the reaction zone. Such reactants, if visible, would appear as a ring around the reaction zone, and the degree of separation is dependent on the volume of solvent zone, and the degree of separation is dependent on the volume of solvent used and the Rf values for the reactants. Small volumes have been found effective for good separation, thus providing a quick, economical, and reliable assay procedure. Typically, solvent volumes of from about 10 μl to about 150 μl, preferably 25 to 100 μl are employed. The solvents may conveniently be applied to the reaction zone with a pipette or hypodermic syringe.
When the present method is being utilized as an enzyme-immunoassay, a solution containing an enzyme substrate may be used for the dual function of applying the substrate and as a chromatographic solvent. Such a procedure reduces the total number of steps and thus reduces the time required to conduct an assay.
After the ligand assay reaction has been completed and the unbound labeled indicator has been separated from the reaction zone, the reaction zone is observed, either visually or with the aid of appropriate instruments, to determine the magnitude of the signal generated by the labeled indicator. This signal might be a measure of radioactivity in the case of a radioimmunoassay, or a colorimetric, ultraviolet or fluorescence response in the case of an enzyme-immunoassay. Typically, colorimetric, ultraviolet, or fluorescent assays may be used for rate determinations, where the rate of formation (or disappearance) of the measured chromophore or fluorophore is compared to a calibration standard as as indication of concentration. Such measurements may be made directly from the porous medium, employing front surface fluorometers or reflectometers. In its simplest form, the present method may be used as a qualitative or semiquantitative test, wherein the reaction zone is observed visually, under visible or ultraviolet light, to determine whether a reaction has occurred or to obtain an approximate indication of the extent of the reaction.
The present invention is further illustrated by the following examples, which are not intended to be limiting.
This example describes a competitive type of enzymeimmunoassay for serum digoxin in accordance with the present invention.
A. Preparation of Anti-Digoxin Papers.
1. Diluent Buffer--1×10-2 M Phosphate, 1.5×10-1 M NaCl, 1×10-2 M EDTA, pH 7.3, with 0.5% bovine serum albumin and 0.1% NaN3.
2. First antibody-Solution--16 microliters of a 1:100 dilution of rabbit anti-digoxin plus 10 microliters of normal rabbit serum was diluted to a final volume of 2000 microliters.
3. Second antibody solution--1:100 dilution of goat anti-rabbit serum.
4. Paper Matrix--Whatman type GF/F glass microfiber filter paper.
To 175 microliters of first antibody solution was added 175 microliters of second antibody solution. The solution was mixed rapidly and immediately pipetted in 50 microliter aliquots onto the paper matrix. The anti-digoxin papers were incubated in a moist condition at 4° C. for 18 hours. They were then washed with 100 microliters of 1.5×10-1 M NaCl containing 0.1% Triton X-100 which was applied to the center of the antibody spot diffusing out radially. Papers were dried under vacuum and stored at 4° C.
B. Preparation of Enzyme Labeled Digoxin.
1. Digoxin--100 milligrams in 10 milliliters of methanol.
2. Periodate--214 milligrams of sodium periodate in 10 milliliters of 100 millimolar potassium phosphate pH 8.0.
3. Alkaline Phosphatase (ALP)--from E. coli dialized against 100 mM Tris buffer pH 8.
4. Sodium Cyanoborohydride--6 milligrams per milliliter in 500 millimolar phosphate pH 6.0.
To 500 microliters of digoxin in methanol was added 500 microliters of periodate suspension. The materials were reacted with agitation for 1 hour in the dark at room temperature. The mixture was centrifuged to remove the periodate. The pH of the supernate was adjusted to 8 with potassium carbonate. The reaction was effective to oxidize the vicinal hydroxyls of the digitose, cleaving the ring and forming the dialdehyde. To the oxidized digoxin was added 200 microliters containing 2 milligrams of alkaline phosphatase. The mixture was incubated in the dark at room temperature for 1 hour. Octanol (1 drop) was added to retard foaming and the pH was adjusted to about 6 with formic acid. The reaction produces Schiff base bonds between the amine groups of the enzyme and the aldehyde groups of the digoxin. The excess aldehydes and the Schiff bases are then reduced by the dropwise addition of 300 microliters of sodium cyanoborohydride. The conjugate thus formed was purified by dialysis (3 times) against saline and by gel filitration chromatography on a Sephadex G-25 column using 10 millimolar Tris, 1.5×10-1 molar NaCl, pH 8.0 as eluant.
C. Preparation of Calibrator-Trace Mixtures
1. The enzyme labeled digoxin was added to commercial normal human serum calibrators, containing azide preservatives, to form calibration-tracer mixtures having a free digoxin concentration of 0 to 6 nanograms per millimeter and a tracer digoxin concentration of 1.6 nanograms per milliliter.
D. Preparation of Substrate Wash Solution
1. A 10 milligram amount of 4-methylumbelliferyl phosphate was added to 100 milliliters of 1.5 molar Tris, 1 millimolar magnesium chloride, pH 8.0.
II. ASSAY OF DIGOXIN
Three anti-digoxin paper spots were labeled for each calibrator-tracer mixture and each sample-tracer mixture. An aliquot of 50 microliters of the appropriate calibrator-tracer mixture or sample-tracer mixture was pipetted onto the center of the anti-digoxin paper spot. The mixture was quickly absorbed into the matrix. The reaction spot was incubated in a moist chamber at room temperature for 5 minutes. An aliquot of 100 microliters of substrate wash solution was applied slowly (60 seconds) to the center of the spot. The wash solution diffused out radially carrying with it any digoxin-ALP not bound by the anti-digoxin immobilized within the matrix. When the diffusion of the substrate begins to slow, the reaction rate of the antibody bound digoxin-ALP is measured in an area about the center of the antibody spot within a radius of 4.5 millimeters.
A delimited area (4.5 millimeter radius) is chosen for the monitoring of the bound enzyme substrate reaction so as to sample a homogenous area of the anti-digoxin reaction matrix (total radius of 6 millimeters). At the same time the free digoxin-ALP has been removed from the monitored area by the subtrate wash solution (13 millimeter radius).
The enzymatic conversion of the non-fluorescent substrate, 4-methylumbelliferyl phosphate, to the fluorescent product, 4-methylumbelliferone, by the bound digoxin-ALP is measured in a suitable front surface fluorometer.
This example describes a competitive type of radioimmunoassay for serum digoxin in accordance with the present invention.
A. Preparation of Anti-Digoxin Papers
1. Diluent Buffer--1×10-2 M phosphate, 1.5×10-1 M NaCl, 1×10-2 M Na2 EDTA, pH 7.3, with 0.5% bovine serum albumin and 0.1% NaN3.
2. Paper Matrix--Toyo type GA-100 glass microfiber filter paper.
To 0.67 milliliters of buffer was added 100 microliters of normal rabbit serum, 20 microliters of 2% patent blue dye and 20 microliters of a 1:100 dilution of anti-digoxin serum. The solution was mixed thoroughly. To this was added 200 microliters of goat anti-rabbit serum. The solution was mixed rapidly and immediately pipetted in 100 microliter aliquots onto the paper matrix. The anti-digoxin papers were incubated in a moist condition at 4° C. for 18 hours. They were then washed with 2 aliquots of 200 microliters each of 100 mM PO4 -saline which was applied to the center of the antibody spot, diffusing out radially. Papers were dried under vacuum and stored at 4° C.
B. Preparation of 125 I Labeled Digoxin.
A commercially available 125 I-digoxin derivative was diluted to 915 picograms per milliliter in 1×10-2 M phosphate, 1.5×10-1 M NaCl, 1×10-2 M EDTA, pH 7.3, with 0.5% bovine serum albumin and 0.1% NaN3.
C. Preparation of Calibrator-Tracer Mixtures.
The 125 I-digoxin solution was added to commercial normal human serum calibrators containing azide preservative, to form calibration-tracer mixtures having a free digoxin concentration of 0 to 6 nanograms per milliliter and a tracer digoxin concentration of 458 picograms per milliliter.
D. Wash Solution.
1×10-2 M phosphate, 1.5×10-1 M NaCl.
II. ASSAY OF DIGOXIN.
Anti-digoxin paper spots were labeled for each calibrator-tracer mixture. An aliquot of 100 microliters of the appropriate calibrator-tracer mixture was pipetted onto the center of the anti-digoxin paper spot. The mixture was quickly absorbed into the matrix. The reaction spot was incubated in open air, on a wire rack at room temperature for 3 minutes. Two aliquots of 200 microliters each of wash solution was applied to the center of the spot. The wash solution diffused out radially carrying with any 125 I-digoxin not bound by the anti-digoxin immobilized within the matrix. A disc (radius 9 millimeters) was then punched from the center of the anti-digoxin paper spot. The discs were counted in a gamma scintillation counter for 1 minute each. The number of counts for each disc was found to be inversely proportional to the concentration of digoxin applied to the disc.
This example describes a sandwich type of radioimmunoassay for serum thyroid stimulating hormone (TSH) in accordance with the present invention.
A. Preparation of anti-TSH papers
1. Diluent Buffer--50 millimolar phosphate, pH 7.4, with 0.1% bovine serum albumin and 0.02% azide.
2. Filter Paper Matrix--Toyo glass fiber paper type GA-100.
To 2.25 milliliters of buffer was added 2.5 milliliters of a 1:100 dilution of goat anti-TSH serum and 250 microliters of rabbit anti-goat serum. The solution was mixed rapidly and immediately pipetted 100 microliter aliquots onto the paper matrix. The spots were incubated in a moist condition at 4° C. for 18 hours. They were then washed 2 times with 200 microliter aliquots of 100 millimolar phosphate buffer pH 7.3. The papers were dried under vacuum and stored at 4° C.
B. 125 I-Anti-TSH Tracer.
A commercial preparation of 125 I labeled rabbit anti-TSH serum containing 58,000 counts per minute per 100 microliters.
C. TSH Calibrators
A commercial secondary standard material was diluted in charcoal treated normal human serum to concentrations of 0, 1.5, 6, 12, 25 and 50 microunits per milliliter.
D. Pre-Application of Tracer to Paper.
To each anti-TSH spot was added 100 microliters of rabbit 125 I-anti-TSH. The spots were dried under vacuum.
II. ASSAY OF TSH.
One anti-TSH paper spot was labeled for each TSH calibrator. 100 microliters of the appropriate calibrator was applied onto the spot. The papers were incubated in a moist chamber for 1 hour at 37° C. Papers were then washed with 2 aliquots of 200 microliters each of 10 mM phosphate pH 7.4. A disc of 9 mm was cut from the center of each spot. All discs were counted in a gamma counter for 1 minute. The number of counts for each disc was found to be proportional to the concentration of TSH applied to the disc.
This example describes a sandwich type of enzyme immunoassay for human IgG in accordance with the present invention.
A. Preparation of Anti-Human IgG Papers
1. Diluent Buffer--0.01M Tris with 0.1% BSA and 0.5% sodium azide, pH 8.0.
2. First Antibody Solution--2 microliters of goat anti-human IgG is diluted to a final volume of 2000 microliters.
3. Second Antibody Solution--1:100 dilution of rabbit anti-goat serum.
4. Paper Matrix--Whatman type GF/F glass microfiber filter paper.
To 175 microliters of first antibody solution is added 175 microliters of second antibody solution. The solution is mixed rapidly and is immediately pipetted in 50 microliter aliquots onto the paper matrix. The anti-human IgG papers are incubated in a moist condition at 4° C. for 18 hours. Papers are stored at 4° C.
B. Preparation of enzyme labeled anti-human IgG--a modification of the method described by A. Murayamd et al. Immuno-chemistry 15: 523 (1978).
1. Anti-human IgG--20 ml of rabbit anti-human IgG serum is mixed for 4 hours at room temperature with 0.4 gms of Aerosil 380. The material is centrifuged and the supernate collected. The treated rabbit anti-human IgG serum is then diluted with equal volume of 0.1M ethylene diamine (EDA) buffer pH 7.0 and applied to a column of QAE Sephadex which has been equilibrated with the EDA buffer. The column is eluted with additional EDA buffer. The protein fractions which are eluted are pooled and concentrated to 20 mg/ml in an Amicon cell fitted with a type PM10 membrane. The material is then dialized against 0.01M K H2 PO4.
2. Periodate--32.5 milligrams of sodium periodate are combined with 177.5 milligrams of KCl and 3.0 ml of methanol.
3. Alkaline Phosphatase (ALP)--from E. coli. The enzyme is dialized against 0.01M phosphate buffer, pH 6.5.
4. Sodium Cyanoborohydride--6 milligrams per milliliter in 0.5M phosphate pH 6.0.
To a test tube wrapped in foil containing 45 μl of purified rabbit anti-human IgG is added 80 μl of periodate suspension. The pH is adjusted to between 4.0 and 4.5 with 1N HCl. The mixture is incubated at room temperature with constant mixing for 30 minutes. The mixture is centrifuged and the supernate collected. 20 μl packed volume of G-25 Sephadex equilibrated with 0.01M K H2 PO4 is added to the supernate. The Sephadex is mixed by inverting for 20 minutes at room temperature. The Sephadex is then centrifuged down and the supernate is collected. 16 mg of ALP is added to the supernate and the pH is adjusted to 6.5 with 0.5M K H2 PO4. The solution is mixed and then dialized against 0.01M phosphate buffer pH 6.5 for 18 hours at room temperature. To the dialized conjugate is added 50 μl sodium cyanoborohydride. The mixture is allowed to react at room temperature for 1 hour. The solution is then dialized against 0.01M Tris 0.15M NaCl pH 8.0.
The conjugate is applied to a Biogel A 1.5 column and the fractions containing both enzyme and antibody activities are collected.
C. Preparation of human IgG calibrators. Human IgG is weighed out and dissolved in 0.01M Tris with 1.0% gelatin and 0.1% sodium azide, pH 8.0 to form calibrators having human IgG concentrations of between 0 mg/ml and 50 mg/ml.
D. Preparation of Substrate Wash Solution. A 10 milligram amount of 4-methylumbelliferyl phosphate is added to 100 milliliters of 1.5M Tris, 0.001M MgCl2, pH 8.0.
II. ASSAY OF HUMAN IgG.
Two anti-human IgG paper spots are labeled for each calibrator and each sample. Calibrators and samples are diluted 1:50 in 0.01M Tris, 0.15M NaCl pH 8.0.
The diluted calibrators and samples are pipetted in 50 microliter aliquots onto the center of the anti-human IgG paper spot. The mixture is quickly absorbed into the matrix. The reaction spot is incubated in a moist chamber at room temperature for 5 minutes and the human IgG in the sample is bound by the antibody immobilized within the matrix. An aliquot of 50 microliters of ALP-anti-human IgG conjugate is applied and is incubated at room temperature for 5 minutes. The conjugate binds to the human IgG bound within the matrix. An aliquot of 100 microliters of substrate wash solution is applied slowly to the center of the spot. The wash solution diffuses out radially carrying with it any ALP-anti-human IgG conjugate not bound within the matrix. When the diffusion of the substrate begins to slow, the reaction rate of the bound ALP-anti-human IgG is measured in a suitable fluorometer.
|Cited Patent||Filing date||Publication date||Applicant||Title|
|US2854317 *||Jun 29, 1956||Sep 30, 1958||Miles Lab||Method and composition for testing bilirubin in urine|
|US2893843 *||Feb 3, 1956||Jul 7, 1959||Miles Lab||Composition of matter|
|US2990253 *||May 21, 1959||Jun 27, 1961||Miles Lab||Diagnostic composition|
|US3061523 *||Nov 7, 1956||Oct 30, 1962||Miles Lab||Method for determining glucose in blood|
|US3645687 *||Jan 5, 1970||Feb 29, 1972||Samuel T Nerenberg||Immunodiffusion plate apparatus|
|US3791932 *||Jan 27, 1972||Feb 12, 1974||Akzona Inc||Process for the demonstration and determination of reaction components having specific binding affinity for each other|
|US3888629 *||Sep 8, 1972||Jun 10, 1975||Kenneth Dawson Bagshawe||Performance of chemical or biological reactions within an absorbent matrix pad|
|US3951748 *||Nov 11, 1974||Apr 20, 1976||Medical Products, Inc.||Sensitized matrix for detection of disease|
|US4046514 *||Nov 24, 1976||Sep 6, 1977||Miles Laboratories, Inc.||Test device and method for determining a component in a sample|
|US4059405 *||Aug 19, 1976||Nov 22, 1977||Damon Corporation||Method and apparatus for analysis of constituent carried in fibrous medium|
|US4071315 *||Jul 12, 1976||Jan 31, 1978||Guy Chateau||Analytical tape medium|
|US4094647 *||Jul 2, 1976||Jun 13, 1978||Thyroid Diagnostics, Inc.||Test device|
|US4157895 *||Oct 7, 1977||Jun 12, 1979||Nuclear International Corporation||RIA reagents and processes|
|US4168146 *||Jan 15, 1976||Sep 18, 1979||Ab Kabi||Immunoassay with test strip having antibodies bound thereto|
|US4258001 *||Dec 27, 1978||Mar 24, 1981||Eastman Kodak Company||Element, structure and method for the analysis or transport of liquids|
|US4299916 *||Dec 26, 1979||Nov 10, 1981||Syva Company||Preferential signal production on a surface in immunoassays|
|US4363874 *||Aug 7, 1981||Dec 14, 1982||Miles Laboratories, Inc.||Multilayer analytical element having an impermeable radiation nondiffusing reflecting layer|
|US4366241 *||Aug 7, 1980||Dec 28, 1982||Syva Company||Concentrating zone method in heterogeneous immunoassays|
|1||Giegel et al., "Radial Partition Immunoassay", Clin. Chem., vol. 28, No. 9, (1982), pp. 1894-1898.|
|2||*||Giegel et al., Radial Partition Immunoassay , Clin. Chem., vol. 28, No. 9, (1982), pp. 1894 1898.|
|Citing Patent||Filing date||Publication date||Applicant||Title|
|US4623461 *||May 31, 1985||Nov 18, 1986||Murex Corporation||Transverse flow diagnostic device|
|US4717654 *||Jun 19, 1984||Jan 5, 1988||Akzo N.V.||Process for solid phase platelet antibody assay|
|US4752562 *||Oct 17, 1984||Jun 21, 1988||Baxter Travenol Laboratories, Inc.||Detection of serum antibody and surface antigen by radial partition immunoassay|
|US4774174 *||Apr 3, 1985||Sep 27, 1988||Baxter Travenol Laboratories, Inc.||Solid phase system for ligand assay|
|US4786606 *||Mar 7, 1985||Nov 22, 1988||Baxter Travenol Laboratories, Inc.||Solid phase system for ligand assay|
|US4806311 *||Aug 28, 1985||Feb 21, 1989||Miles Inc.||Multizone analytical element having labeled reagent concentration zone|
|US4806312 *||Aug 28, 1985||Feb 21, 1989||Miles Inc.||Multizone analytical element having detectable signal concentrating zone|
|US4855240 *||May 13, 1987||Aug 8, 1989||Becton Dickinson And Company||Solid phase assay employing capillary flow|
|US4857453 *||Apr 7, 1987||Aug 15, 1989||Syntex (U.S.A.) Inc.||Immunoassay device|
|US4857471 *||Jul 20, 1987||Aug 15, 1989||Eastman Kodak Company||Analyzer with wash station separate from incubator|
|US4948726 *||Mar 20, 1989||Aug 14, 1990||Longoria Claude C||Enzyme immunoassay based on membrane separation of antigen-antibody complexes|
|US4960691 *||Sep 29, 1986||Oct 2, 1990||Abbott Laboratories||Chromatographic test strip for determining ligands or receptors|
|US4999285 *||Nov 15, 1984||Mar 12, 1991||Syntex (U.S.A.) Inc.||Chromatographic cassette|
|US5063151 *||Sep 20, 1985||Nov 5, 1991||Biometallics, Inc.||Immunoassay method and kit|
|US5073484 *||Feb 23, 1983||Dec 17, 1991||Bio-Metric Systems, Inc.||Quantitative analysis apparatus and method|
|US5120643 *||Jul 13, 1987||Jun 9, 1992||Abbott Laboratories||Process for immunochromatography with colloidal particles|
|US5147609 *||May 19, 1989||Sep 15, 1992||Pb Diagnostic Systems, Inc.||Assay element|
|US5149622 *||May 21, 1990||Sep 22, 1992||Abbott Laboratories||Solid phase analytical device and method for using same|
|US5176999 *||Dec 7, 1989||Jan 5, 1993||Eastman Kodak Company||Buffered wash composition, insolubilizing composition, test kits and method of use|
|US5212065 *||Oct 25, 1990||May 18, 1993||International Diagnostic Systems, Corp.||Rapid assay device|
|US5256372 *||Dec 5, 1989||Oct 26, 1993||Idexx Corporation||Dipstick test device including a removable filter assembly|
|US5354654 *||Jul 16, 1993||Oct 11, 1994||The United States Of America As Represented By The Secretary Of The Navy||Lyophilized ligand-receptor complexes for assays and sensors|
|US5366864 *||Sep 30, 1992||Nov 22, 1994||Eastman Kodak Company||Buffered wash composition, insolubilizing composition, test kits and method of use|
|US5376556 *||Sep 11, 1992||Dec 27, 1994||Abbott Laboratories||Surface-enhanced Raman spectroscopy immunoassay|
|US5434087 *||Feb 24, 1993||Jul 18, 1995||Abbott Laboratories||Folate immunoassay utilizing folate binding protein in a multiclonal antibody format|
|US5459078 *||Jun 9, 1993||Oct 17, 1995||Abbott Laboratories||Methods and reagents for performing ion-capture digoxin assays|
|US5459080 *||Jan 27, 1994||Oct 17, 1995||Abbott Laboratories||Ion-capture assays using a specific binding member conjugated to carboxymethylamylose|
|US5468622 *||Jul 26, 1993||Nov 21, 1995||Immunomatrix, Inc.||Salt stabilization of antibody-enzyme conjugates heat-dried into paper|
|US5468648 *||Dec 7, 1993||Nov 21, 1995||Smithkline Diagnostics, Inc.||Interrupted-flow assay device|
|US5525514 *||Apr 3, 1995||Jun 11, 1996||Johnson & Johnson Clinical Diagnostics, Inc.||Wash detection method for dried chemistry test elements|
|US5578577 *||Jun 6, 1995||Nov 26, 1996||Abbott Laboratories||Method for storing labile proteins|
|US5607863 *||Dec 7, 1993||Mar 4, 1997||Smithkline Diagnostics, Inc.||Barrier-controlled assay device|
|US5620860 *||Feb 24, 1995||Apr 15, 1997||Johnson & Johnson Clinical Diagnostics||Method for washing immunoassay elements|
|US5622871 *||Jul 15, 1993||Apr 22, 1997||Unilever Patent Holdings B.V.||Capillary immunoassay and device therefor comprising mobilizable particulate labelled reagents|
|US5637468 *||Sep 21, 1995||Jun 10, 1997||Immunomatrix, Inc.||Salt stabilization of antibody-enzyme conjugates heat-dried into paper|
|US5641688 *||Jun 6, 1995||Jun 24, 1997||Johnson & Johnson Clinical Diagnostics, Inc.||Immunoassay including washing a slide at different locations|
|US5654162 *||Jun 1, 1992||Aug 5, 1997||Bio-Metric Systems, Inc.||Chemical analysis apparatus and method|
|US5656503 *||Sep 15, 1994||Aug 12, 1997||Unilever Patent Holdings B.V.||Test device for detecting analytes in biological samples|
|US5660990 *||Aug 18, 1995||Aug 26, 1997||Immunivest Corporation||Surface immobilization of magnetically collected materials|
|US5714340 *||Jun 16, 1994||Feb 3, 1998||Johnson & Johnson Clinical Diagnostics, Inc.||Immunoassay elements having a receptor zone|
|US5741714 *||Jul 16, 1996||Apr 21, 1998||Immunivest Corporation||Detection of bound analyte by magnetic partitioning and masking|
|US5753519 *||Jun 6, 1995||May 19, 1998||Cornell Research Foundation, Inc.||Liposome-enhanced immunoaggregation assay and test device|
|US5756362 *||Feb 1, 1995||May 26, 1998||Cornell Research Foundation, Inc.||Liposome-enhanced immunoaggregation assay and test device|
|US5789154 *||Oct 12, 1993||Aug 4, 1998||Cornell Research Foundation, Inc.||Liposome-enhanced immunoassay and test device|
|US5795784||Sep 19, 1996||Aug 18, 1998||Abbott Laboratories||Method of performing a process for determining an item of interest in a sample|
|US5824268 *||Oct 14, 1997||Oct 20, 1998||Universal Health Watch, Inc.||Rapid self-contained assay format|
|US5837546 *||Jun 7, 1996||Nov 17, 1998||Metrika, Inc.||Electronic assay device and method|
|US5846838 *||Jun 18, 1997||Dec 8, 1998||Smithkline Diagnostics, Inc.||Opposable-element assay device employing conductive barrier|
|US5851777 *||Feb 5, 1996||Dec 22, 1998||Dade Behring Inc.||Homogeneous sol-sol assay|
|US5856194||Sep 19, 1996||Jan 5, 1999||Abbott Laboratories||Method for determination of item of interest in a sample|
|US5861319 *||Oct 28, 1994||Jan 19, 1999||Dade Behring Inc.||Immobilization of specific binding assay reagents|
|US5866322 *||Oct 11, 1991||Feb 2, 1999||Abbott Laboratories||Method for performing Rubella assay|
|US5869345 *||Jun 2, 1995||Feb 9, 1999||Smithkline Diagnostics, Inc.||Opposable-element assay device employing conductive barrier|
|US5877028||Mar 31, 1993||Mar 2, 1999||Smithkline Diagnostics, Inc.||Immunochromatographic assay device|
|US5879951 *||Jan 29, 1997||Mar 9, 1999||Smithkline Diagnostics, Inc.||Opposable-element assay device employing unidirectional flow|
|US5898005 *||Apr 13, 1994||Apr 27, 1999||Dade Behring Inc.||Rapid detection of analytes with receptors immobilized on soluble submicron particles|
|US5939252 *||May 9, 1997||Aug 17, 1999||Lennon; Donald J.||Detachable-element assay device|
|US5945345 *||Aug 27, 1996||Aug 31, 1999||Metrika, Inc.||Device for preventing assay interference using silver or lead to remove the interferant|
|US5948630 *||Aug 5, 1996||Sep 7, 1999||Dade Behring Inc.||Method and composition for reducing the effects of endogenous alkaline phosphatase|
|US5958339 *||Aug 31, 1992||Sep 28, 1999||Clinical Diagnostic Systems, Inc.||Format for immunoassay in thin film|
|US5958790 *||May 22, 1995||Sep 28, 1999||Nycomed Imaging As||Solid phase transverse diffusion assay|
|US5968839 *||May 13, 1996||Oct 19, 1999||Metrika, Inc.||Method and device producing a predetermined distribution of detectable change in assays|
|US5998220||Feb 10, 1994||Dec 7, 1999||Beckman Coulter, Inc.||Opposable-element assay devices, kits, and methods employing them|
|US6017767||Jun 5, 1995||Jan 25, 2000||Beckman Coulter, Inc.||Assay device|
|US6020147 *||Jun 1, 1992||Feb 1, 2000||Surmodics, Inc.||Chemical analysis apparatus and method|
|US6083708 *||Aug 11, 1995||Jul 4, 2000||Dade Behring Inc.||Polypeptide: dendrimer complexes|
|US6086748 *||Feb 20, 1998||Jul 11, 2000||Cornell Research Foundation, Inc.||Liposome enhanced immunoaggregation assay and test device|
|US6100099 *||Oct 28, 1998||Aug 8, 2000||Abbott Laboratories||Test strip having a diagonal array of capture spots|
|US6121056 *||Apr 12, 1994||Sep 19, 2000||Dade Behring Inc.||Random detection of antigens with antibodies immobilized on soluble submicron particles|
|US6168956||May 29, 1991||Jan 2, 2001||Beckman Coulter, Inc.||Multiple component chromatographic assay device|
|US6187598||Jun 7, 1995||Feb 13, 2001||Conopco Inc.||Capillary immunoassay and device therefor comprising mobilizable particulate labelled reagents|
|US6210898||Sep 16, 1999||Apr 3, 2001||Abbott Laboratories||Method of performing immunochromatography|
|US6228660||Jun 7, 1995||May 8, 2001||Conopco Inc.||Capillary immunoassay and device therefor comprising mobilizable particulate labelled reagents|
|US6248596||Mar 3, 1998||Jun 19, 2001||Cornell Research Foundation, Inc.||Liposome-enhanced immunoassay and test device|
|US6271044 *||May 6, 1998||Aug 7, 2001||University Of Pittsburgh Of The Commonwealth System Of Higher Education||Method and kit for detecting an analyte|
|US6297060||Feb 11, 1999||Oct 2, 2001||Biosite Diagnostics, Inc.||Assay devices comprising a porous capture membrane in fluid-withdrawing contact with a nonabsorbent capillary network|
|US6352862||Jun 9, 1997||Mar 5, 2002||Unilever Patent Holdings B.V.||Analytical test device for imuno assays and methods of using same|
|US6368876||Jan 9, 1998||Apr 9, 2002||Genzyme Diagnostics||One step immunochromatographic device and method of use|
|US6383766||Sep 30, 1999||May 7, 2002||Ortho-Clinical Diagnostics, Inc.||Reduced cortisol conjugates|
|US6534320||Mar 23, 1998||Mar 18, 2003||Abbott Laboratories||Process for immunochromatography with colloidal particles|
|US6562298||Apr 23, 1999||May 13, 2003||Abbott Laboratories||Structure for determination of item of interest in a sample|
|US6576460||Oct 27, 2000||Jun 10, 2003||Cornell Research Foundation, Inc.||Filtration-detection device and method of use|
|US6723840||Nov 28, 2000||Apr 20, 2004||The Regents Of The University Of California||Identification and expression of a novel kinesin motor protein|
|US6764830||Jan 22, 1999||Jul 20, 2004||The Regents Of The University Of California||Thermomyces lanuginosus kinesin motor protein and methods of screening for modulators of kinesin proteins|
|US6815169||Nov 28, 2000||Nov 9, 2004||The Regents Of The University Of California||Identification and expression of a novel kinesin motor protein|
|US6818455||Feb 28, 2001||Nov 16, 2004||Inverness Medical Switzerland Gmbh||Capillary immunoassay and device therefor comprising mobilizable particulate labelled reagents|
|US6924153||Mar 6, 1997||Aug 2, 2005||Quidel Corporation||Quantitative lateral flow assays and devices|
|US6960457||Sep 3, 1998||Nov 1, 2005||Stanford University||Reversible immobilization of arginine-tagged moieties on a silicate surface|
|US7109042||Feb 12, 2001||Sep 19, 2006||Inverness Medical Switzerland Gmbh||Assays|
|US7144742||Dec 17, 2004||Dec 5, 2006||Quidel Corporation||Quantitative lateral flow assays and devices|
|US7150995||Jan 16, 2004||Dec 19, 2006||Metrika, Inc.||Methods and systems for point of care bodily fluid analysis|
|US7157551||Feb 13, 2004||Jan 2, 2007||Cephalon, Inc.||Compositions and methods for the detection and treatment of methylthioadenosine phosphorylase deficient cancers|
|US7186521||Sep 25, 2001||Mar 6, 2007||The Regents Of The University Of California||Determining the effect of a substance on sequestration, uptake, and accumulation of amyloid in brain cells|
|US7238537||Sep 4, 2001||Jul 3, 2007||Inverness Medical Switzerland Gmbh||Assays|
|US7270959||Jul 25, 2001||Sep 18, 2007||Oakville Hong Kong Company Limited||Specimen collection container|
|US7285426||May 5, 2005||Oct 23, 2007||Medtox Scientific, Inc.||Bulking materials containing starch reagent for use in test devices|
|US7297554||Oct 28, 2003||Nov 20, 2007||Microdiagnostics, Inc.||Immunoassay system|
|US7300633||Aug 2, 2002||Nov 27, 2007||Oakville Hong Kong Company Limited||Specimen collection container|
|US7319032||Sep 21, 2004||Jan 15, 2008||Medtox||Non-sugar sweeteners for use in test devices|
|US7321065||Apr 16, 2004||Jan 22, 2008||The Regents Of The University Of California||Thyronamine derivatives and analogs and methods of use thereof|
|US7355079||Apr 5, 2005||Apr 8, 2008||The Regents Of The University Of California||Thyronamine derivatives and analogs and methods of use thereof|
|US7384796||Dec 23, 2002||Jun 10, 2008||Inverness Medical Switzerland Gmbh||Assays|
|US7407813||Dec 23, 2002||Aug 5, 2008||Inverness Medical Switzerland Gmbh||Assays|
|US7419796||Oct 2, 2002||Sep 2, 2008||Cornell Research Foundation||Method of making a test device|
|US7491551||Nov 8, 2006||Feb 17, 2009||Quidel Corporation||Quantitative lateral flow assays and devices|
|US7510858||Aug 2, 2005||Mar 31, 2009||Janssen Pharmaceutica N.V.||IRAK 1c splice variant and its use|
|US7517495||Jan 27, 2004||Apr 14, 2009||Inverness Medical Switzerland Gmbh||Biological specimen collection and analysis system|
|US7531362||Jun 6, 2002||May 12, 2009||Medmira Inc.||Rapid diagnostic assay|
|US7560272||Jan 4, 2003||Jul 14, 2009||Inverness Medical Switzerland Gmbh||Specimen collection and assay container|
|US7598080||Aug 9, 2005||Oct 6, 2009||Carl Deirmengian||Diagnostic assay for source of inflammation|
|US7611858||Apr 20, 2007||Nov 3, 2009||University Of Florida Research Foundation, Inc.||Detection of cannabinoid receptor biomarkers and uses thereof|
|US7635597||Apr 19, 2004||Dec 22, 2009||Bayer Healthcare Llc||Dry reagent particle assay and device having multiple test zones and method therefor|
|US7645584||Jan 12, 2010||University Of Florida Research Foundation, Inc.||Biomarkers of liver injury|
|US7718388||Nov 10, 2006||May 18, 2010||Cornell Research Foundation, Inc.||Universal biosensor and methods of use|
|US7754464||Jul 13, 2010||Pharmacia And Upjohn Company, Llc||Cellular permissivity factor for viruses and uses thereof|
|US7858397||May 28, 2008||Dec 28, 2010||Cornell Research Foundation, Inc.||Dehydration/rehydration of derivatized, marker-loaded liposomes on a test device and method of use thereof|
|US7888470||Jul 17, 2007||Feb 15, 2011||Senomyx, Inc.||Chimeric T1R taste receptor polypeptides and nucleic acid sequences encoding and cell lines that express said chimeric T1R polypeptides|
|US7893197||Aug 25, 2005||Feb 22, 2011||Janssen Pharmaceutica N.V.||Relaxin-3 chimeric polypeptides and their preparation and use|
|US8025850||Sep 27, 2011||Medmira Inc.||Rapid diagnostic device, assay and multifunctional buffer|
|US8048638||Nov 1, 2011||University Of Florida Research Foundation, Inc.||Biomarkers of liver injury|
|US8058050||May 6, 2010||Nov 15, 2011||Pharmacia And Upjohn Company Llc||Cellular permissivity factor for viruses and uses thereof|
|US8193328||Sep 7, 2006||Jun 5, 2012||Philadelphia Health & Education Corporation||Identification of modulators of serine protease inhibitor Kazal and their use as anti-cancer and anti-viral agents|
|US8287817||Oct 16, 2012||Medmira, Inc.||Rapid diagnostic device, assay and multifunctional buffer|
|US8383580||Jun 10, 2011||Feb 26, 2013||The Regents Of The University Of Michigan||Methods of administering lipocalins to treat metabolic disorders and cardiovascular diseases|
|US8394626||Mar 12, 2013||Alere Switzerland Gmbh||Specimen collection and assay container|
|US8404455||Mar 26, 2013||Senomyx, Inc.||Chimeric T1R taste receptor polypeptides and nucleic acid sequences encoding and cell lines that express said chimeric T1R polypeptides|
|US8415455||Sep 3, 2008||Apr 9, 2013||Compugen Ltd||Polypeptides and polynucleotides, and uses thereof as a drug target for producing drugs and biologics|
|US8486685||Sep 29, 2011||Jul 16, 2013||Zoetis P&U Llc||Cellular permissivity factor for viruses and uses thereof|
|US8486717||Jan 4, 2012||Jul 16, 2013||Symbolics, Llc||Lateral flow assays using two dimensional features|
|US8586375||Sep 11, 2012||Nov 19, 2013||Medmira Inc.||Rapid diagnostic device, assay and multifunctional buffer|
|US8652416 *||Oct 15, 2009||Feb 18, 2014||Samsung Electronics Co., Ltd.||Article for assaying target, comprising solid surface on which first binding member, blocking material, and second binding member are immobilized, and use thereof|
|US8735081||Mar 12, 2012||May 27, 2014||Senomyx, Inc.||T1R hetero-oligomeric taste receptors, cell lines that express said receptors, and taste compounds|
|US8865458||Feb 25, 2013||Oct 21, 2014||Alere Switzerland Gmbh||Specimen collection and assay container|
|US8895050||Dec 23, 2011||Nov 25, 2014||Senomyx, Inc.||Flavors, flavor modifiers, tastants, taste enhancers, umami or sweet tastants, and/or enhancers and use thereof|
|US8993231||Mar 17, 2009||Mar 31, 2015||Marshall University Research Corporation||Methods for stem cell production and therapy|
|US9072313||Oct 5, 2012||Jul 7, 2015||Senomyx, Inc.||Comestible compositions comprising high potency savory flavorants, and processes for producing them|
|US9091686||Feb 19, 2013||Jul 28, 2015||Senomyx, Inc.||Chimeric T1R taste receptor polypeptides and nucleic acid sequences encoding and cell lines that express said chimeric T1R polypeptides|
|US9102912||Jul 12, 2013||Aug 11, 2015||Zoetis Services Llc||Cellular permissivity factor for viruses and uses thereof|
|US9107862||Feb 15, 2013||Aug 18, 2015||Compugen Ltd.||Polypeptides and polynucleotides, and uses thereof as a drug target for producing drugs and biologics|
|US9164087||Oct 28, 2013||Oct 20, 2015||Medmira Inc.||Rapid diagnostic device, assay and multifunctional buffer|
|US9216412||Mar 22, 2012||Dec 22, 2015||Cyvek, Inc.||Microfluidic devices and methods of manufacture and use|
|US9229001||Nov 23, 2010||Jan 5, 2016||Cyvek, Inc.||Method and apparatus for performing assays|
|US9316641||Jan 11, 2011||Apr 19, 2016||Biogen Ma Inc.||Assay for JC virus antibodies|
|US20020061515 *||Sep 25, 2001||May 23, 2002||The Regents Of The University Of California||Model for neurodegenerative diseases involving amyloid accumulation|
|US20020106705 *||Aug 21, 2001||Aug 8, 2002||Tularik Inc.||Detection method for monitoring beta tubulin isotype specific modification|
|US20020132370 *||Feb 23, 2001||Sep 19, 2002||Lassen Michael Rud||Detection of a blood coagulation activity marker in a body fluid sample|
|US20030025985 *||Jul 23, 2002||Feb 6, 2003||Seiko Epson Corporation||Electro-optical device, electronic apparatus, method for forming a colored layer, and method for manufacturing the electro-optical device|
|US20030049857 *||Jun 6, 2002||Mar 13, 2003||Chan Hermes K.W.||Rapid diagnostic device, assay and multifunctional buffer|
|US20030143755 *||Dec 23, 2002||Jul 31, 2003||Davis Paul James||Assays|
|US20030219908 *||Dec 23, 2002||Nov 27, 2003||Davis Paul James||Assays|
|US20030228639 *||Mar 19, 2001||Dec 11, 2003||Wright George L||Prostate cancer markers|
|US20040087036 *||Oct 28, 2003||May 6, 2004||Roy Chung||Immunoassay system|
|US20040157249 *||Dec 22, 2003||Aug 12, 2004||Yizhong Gu||Canine L-PBE sequences|
|US20040157263 *||Jun 25, 2003||Aug 12, 2004||Bayer Aktiengesellschaft,||Method for impedimetric detection of one or more analytes in a sample, and device for use therin|
|US20040157295 *||Dec 22, 2003||Aug 12, 2004||Yizhong Gu||Canine dioxin/aryl hydrocarbon receptor sequences|
|US20040158043 *||Dec 22, 2003||Aug 12, 2004||Yizhong Gu||Canine CYP1A2 sequences|
|US20040247600 *||Feb 13, 2004||Dec 9, 2004||Leoni Lorenzo M.||Compositions and methods for the detection and treatment of methylthioadenosine phosphorylase deficient cancers|
|US20050014734 *||Jul 18, 2003||Jan 20, 2005||Genovate Biotechnology Co., Ltd.||Modulation of interleukin-10 by DHEA|
|US20050025779 *||Jun 10, 2004||Feb 3, 2005||Vaxgen, Inc.||HIV-1 envelope glycoproteins having unusual disulfide structure|
|US20050079484 *||Oct 10, 2003||Apr 14, 2005||Heineman William Richard||Method of detecting biological materials in liquid|
|US20050096485 *||Apr 16, 2004||May 5, 2005||Scanlan Thomas S.||Thyronamine derivatives and analogs and methods of use thereof|
|US20050101032 *||Aug 27, 2004||May 12, 2005||Metrika, Inc.||Assay device, composition, and method of optimizing assay sensitivity|
|US20050158866 *||Jan 16, 2004||Jul 21, 2005||Xie Zongcen C.||Methods and systems for point of care bodily fluid analysis|
|US20050170527 *||Dec 17, 2004||Aug 4, 2005||Hans Boehringer||Quantitative lateral flow assays and devices|
|US20050227370 *||Jan 21, 2005||Oct 13, 2005||Ramel Urs A||Body fluid analyte meter & cartridge system for performing combined general chemical and specific binding assays|
|US20050239214 *||Sep 21, 2004||Oct 27, 2005||Medtox Scientific, Inc.||Non-sugar sweeteners for use in test devices|
|US20050255608 *||May 5, 2005||Nov 17, 2005||Medtox Scientific, Inc.||Bulking materials containing starch reagent for use in test devices|
|US20050271685 *||Apr 25, 2005||Dec 8, 2005||Pharmacia And Upjohn Company, Llc||Cellular permissivity factor for viruses and uses thereof|
|US20050283003 *||May 19, 2004||Dec 22, 2005||Spudich James A||Reversible immobilization of arginine-tagged moieties on a silicate surface|
|US20060019404 *||Feb 18, 2005||Jan 26, 2006||Blatt Joel M||Quantitative assay with extended dynamic range|
|US20060035980 *||Apr 5, 2005||Feb 16, 2006||Scanlan Thomas S||Thyronamine derivatives and analogs and methods of use thereof|
|US20060088830 *||Feb 20, 2003||Apr 27, 2006||Eastern Virginia Medical School||Protein biomarkers that distinguish prostate cancer from non-malignant cells|
|US20060246489 *||Mar 31, 2006||Nov 2, 2006||Svetlov Stanislav I||Biomarkers of liver injury|
|US20070104709 *||Aug 6, 2004||May 10, 2007||Xiaodong Li||T1r hetero-oligomeric taste receptors, cell lines that express said receptors, and taste compounds|
|US20070243181 *||Aug 2, 2005||Oct 18, 2007||Wai-Ping Leung||Irak1c Splice Variant And Its Use|
|US20070243630 *||Nov 8, 2006||Oct 18, 2007||Quidel Corporation||Quantitative lateral flow assays and devices|
|US20080051336 *||Aug 25, 2005||Feb 28, 2008||Pascal Bonaventure||Relaxin-3 Chimeric Polypeptides and Their Preparation and Use|
|US20080230383 *||May 28, 2008||Sep 25, 2008||Durst Richard A||Dehydration/rehydration of derivatized, marker-loaded liposomes on a test device and method of use thereof|
|US20090017457 *||Sep 7, 2006||Jan 15, 2009||Xuanyong Lu||Identification of modulators of serine protease inhibitor kazal and their use as anti-cancer and anti-viral agents|
|US20090105347 *||Nov 30, 2007||Apr 23, 2009||The Regents Of The University Of California||Thyronamine derivatives and analogs and methods of use thereof|
|US20090155850 *||Oct 27, 2006||Jun 18, 2009||The Florida International University Board Of Trustees||Horse:Human Chimeric Antibodies|
|US20090318301 *||Aug 28, 2009||Dec 24, 2009||Carl Deirmengian||Diagnostic assay for source of inflammation|
|US20100021472 *||Jul 27, 2009||Jan 28, 2010||Geetha Srikrishna||Methods for diagnosing and treating cancer|
|US20100034699 *||Feb 11, 2010||Medmira Inc.||Rapid diagnostic device, assay and multifunctional buffer|
|US20100062435 *||Mar 17, 2009||Mar 11, 2010||Marshall University Research Corporation||Methods for Stem Cell Production and Therapy|
|US20100105071 *||Feb 27, 2008||Apr 29, 2010||Children's Medical Center Corporation||Methods for predicting the onset of menarche|
|US20100159483 *||Oct 15, 2009||Jun 24, 2010||Samsung Electronics Co., Ltd.||Article for assaying target, comprising solid surface on which first binding member, blocking material, and second binding member are immobilized, and use thereof|
|US20100196942 *||Jan 12, 2010||Aug 5, 2010||University Of Florida Research Foundation, Inc.||Biomarkers of liver injury|
|US20100291100 *||Mar 26, 2010||Nov 18, 2010||Gojo Industries, Inc.||Compositions And Methods For Screening And Using Compounds Antagonizing Spore-Surface Interactions|
|US20110052501 *||Feb 1, 2009||Mar 3, 2011||Liat Dassa||Polypeptides and polynucleotides, and uses thereof as a drug target for producing drugs and biologics|
|US20110091454 *||Feb 8, 2010||Apr 21, 2011||Alex Diber||Methods and systems for annotating biomolecular sequences|
|US20110136166 *||Mar 13, 2009||Jun 9, 2011||Eastern Virginia Medical School||Imaging Mass Spectrometry for Improved Prostrate Cancer Diagnostics|
|US20110143378 *||Jun 16, 2011||CyVek LLC.||Microfluidic method and apparatus for high performance biological assays|
|US20110184747 *||Aug 9, 2007||Jul 28, 2011||Carmen Bozic||Method for distribution of a drug|
|USRE34405 *||Nov 12, 1987||Oct 12, 1993||Abbott Laboratories||Determination of analytes in particle-containing medium|
|USRE37437 *||Oct 18, 1995||Nov 6, 2001||Dade Behring Marburg, Gmbh||Sheet-like diagnostic device|
|USRE38688 *||Mar 30, 2001||Jan 18, 2005||Dade Behring Marburg Gmbh||Sheet-like diagnostic device|
|EP0232816A2 *||Jan 30, 1987||Aug 19, 1987||Miles Inc.||Heterogeneous specific binding assay employing an aggregatable binding reagent|
|EP0675361A2||Jul 12, 1988||Oct 4, 1995||Abbott Laboratories||Method for storage of labile protein materials|
|EP0729029A2 *||Feb 23, 1996||Aug 28, 1996||JOHNSON & JOHNSON CLINICAL DIAGNOSTICS, INC.||A method for washing immunoassay elements|
|EP0747702A2 *||Jun 5, 1996||Dec 11, 1996||JOHNSON & JOHNSON CLINICAL DIAGNOSTICS, INC.||Method for washing immunoassay elements|
|EP1333094A2||Oct 1, 1997||Aug 6, 2003||Geron Corporation||Human telomerase catalytic subunit|
|EP1783139A2||Oct 1, 1997||May 9, 2007||Geron Corporation||Human telomerase catalytic subunit|
|EP1953556A2||Aug 11, 2003||Aug 6, 2008||Bayer HealthCare AG||Polypeptide biomarkers for diagnosing Alzheimer's disease|
|EP1961293A2||Sep 24, 2002||Aug 27, 2008||Pioneer Hi-Bred International, Inc.||Phytate polynucleotides and methods of use|
|EP1985630A2||Jan 3, 2002||Oct 29, 2008||Senomyx, Inc.||T1R taste receptors and genes encoding same|
|EP2003451A2||Jun 26, 2002||Dec 17, 2008||Senomyx, Inc.||T1R hetero-oligomeric taste receptors and cell lines that express said receptors and use thereof for identification of taste compounds|
|EP2026073A1||Apr 30, 2001||Feb 18, 2009||University Of Iowa Research Foundation||Diagnostics and therapeutics for macular degeneration-related disorders|
|EP2113566A1||Jul 15, 1997||Nov 4, 2009||The Regents Of The University Of California||Genes from 20q13 amplicon and their uses|
|EP2153848A2||Jan 26, 2006||Feb 17, 2010||The Regents of the University of California||Therapeutic monoclonal antibodies that neutralize botulinium neurotoxins|
|EP2172478A2||Feb 6, 1998||Apr 7, 2010||THE GOVERNMENT OF THE UNITED STATES OF AMERICA as represented by THE SECRETARY OF THE DEPARTMENT OF HEALTH AND HUMAN SERVICES||Activity-dependent neurotrophic factor III (ADNF III)|
|EP2204385A1||Nov 24, 2004||Jul 7, 2010||The Government Of U.S.A, As Represented By Secretary, Department of Health and Human Sevices||Pseudomonas exotoxin A mutants and uses thereof|
|EP2207033A2||Apr 15, 2005||Jul 14, 2010||University of Florida Research Foundation, Inc.||Neural proteins as biomarkers for nervous system injury and other neural disorders|
|EP2213740A1||Oct 1, 1997||Aug 4, 2010||The Regents of the University of Colorado||Human telomerase catalytic subunit|
|EP2216339A1||Jan 16, 2007||Aug 11, 2010||Compugen Ltd.||Novel nucleotide and amino acid sequences, and methods of use thereof for diagnosis|
|EP2221620A2||Aug 11, 2003||Aug 25, 2010||Bayer Schering Pharma AG||Polypeptide biomarkers for diagnosing Alzheimer's disease|
|EP2258726A1||Jun 13, 1996||Dec 8, 2010||The Regents of the University of California||High affinity human antibodies to c-erbB-2|
|EP2292795A2||Oct 21, 2004||Mar 9, 2011||Orion Genomics, LLC||Differential enzymatic fragmatation|
|EP2293067A2||Jun 26, 2002||Mar 9, 2011||Senomyx, Inc.||T1R hetero-oligomeric taste receptors and cell lines that express said receptors and use thereof for identification of taste compounds|
|EP2298802A1||Mar 7, 2001||Mar 23, 2011||Senomyx, Inc.||T1R3 taste receptors and genes encoding same|
|EP2327985A2||Jun 26, 2002||Jun 1, 2011||Senomyx, Inc.||T1R Hetero-oligomeric taste receptors and cell lines that express said receptors and use thereof for identification of taste compounds|
|EP2381256A1||Jul 3, 2002||Oct 26, 2011||The Regents of The University of California||Mammalian sweet and amino acid heterodimeric taste receptors|
|EP2386571A2||Apr 10, 2006||Nov 16, 2011||BioGeneriX AG||Compositions and methods for the preparation of protease resistant human growth hormone glycosylation mutants|
|EP2392925A1||Jun 27, 2008||Dec 7, 2011||Abbott Laboratories||Methods and marker combinations for screening for predisposition to lung cancer|
|EP2436774A2||Aug 1, 2003||Apr 4, 2012||The Regents of The University of California||Therapeutic monoclonal antibodies that neutralize botulinum neurotoxins|
|EP2453024A2||Jun 21, 2005||May 16, 2012||The Board of Trustees of The Leland Stanford Junior University||Genes and pathways differentially expressed in bipolar disorder and/or major depressive disorder|
|EP2477032A1||Jan 18, 2011||Jul 18, 2012||Baxter International Inc||Measurement of anti-amyloid antibodies in human blood|
|EP2485047A2||Dec 21, 2006||Aug 8, 2012||Abbott Laboratories||Methods and marker combinations for screening for predisposition to lung cancer|
|EP2520310A1||Apr 5, 2005||Nov 7, 2012||Pharmacia & Upjohn Company LLC||Cellular permissivity factor for viruses, and uses thereof|
|EP2564864A2||Nov 13, 2006||Mar 6, 2013||The Board of Trustees of the Leland||FGF2-related methods for diagnosing and treating depression|
|EP2567970A1||Oct 3, 2006||Mar 13, 2013||Compugen Ltd.||Novel nucleotide and amino acid sequences, and assays and methods of use thereof for diagnosis|
|EP2586456A1||Oct 31, 2005||May 1, 2013||BioGeneriX AG||Remodeling and glycopegylation of fibroblast growth factor (FGF)|
|EP2650683A1||Jun 27, 2008||Oct 16, 2013||Abbott Laboratories||Methods and marker combinations for screening for predisposition to lung cancer|
|EP2712617A1||Mar 14, 2005||Apr 2, 2014||Intercept Pharmaceuticals, Inc.||Treatment of fibrosis using Fxr ligands|
|EP2769728A1||Sep 3, 2008||Aug 27, 2014||Compugen Ltd.|
|EP2769729A1||Sep 3, 2008||Aug 27, 2014||Compugen Ltd.|
|EP2865689A1||Dec 8, 2009||Apr 29, 2015||Compugen Ltd.||FAM26F polypeptides and polynucleotides, and uses thereof as a drug target for producing drugs and biologics|
|EP2918601A1||Oct 3, 2006||Sep 16, 2015||Compugen Ltd.||Novel nucleotide and amino acid sequences, and assays and methods of use thereof for diagnosis|
|WO1987001811A1 *||Sep 19, 1986||Mar 26, 1987||Biometallics Inc.||Immunoassay method and kit|
|WO1988003650A1 *||Nov 9, 1987||May 19, 1988||Gene Labs, Inc.||Blood affinity diagnostic method|
|WO1994019693A1 *||Feb 9, 1994||Sep 1, 1994||Baxter Diagnostics Inc.||Immobilization of specific binding assay reagents|
|WO1995008771A1 *||Sep 7, 1994||Mar 30, 1995||Baxter Diagnostics Inc.||Immobilization of chemically cross-linked proteins on solid supports|
|WO1998014593A2||Oct 1, 1997||Apr 9, 1998||Geron Corporation||Human telomerase catalytic subunit|
|WO1998022800A1 *||Nov 18, 1997||May 28, 1998||Wyntek Diagnostics, Inc.||Hybrid one-step immunochromatographic device and method of use|
|WO2003045335A2||Nov 27, 2002||Jun 5, 2003||Stemcyte, Inc.||Stem cell screening and transplantation therapy for hiv infection|
|WO2005042704A2||Oct 21, 2004||May 12, 2005||Orion Genomics Llc||Differential enzymatic fragmentation|
|WO2005107796A2||Apr 5, 2005||Nov 17, 2005||Pharmacia & Upjohn Company, Llc||Cellular permissivity factor for viruses, and uses thereof|
|WO2007056352A2||Nov 6, 2006||May 18, 2007||The Scripps Research Institute||Compositions and methods for controlling tissue factor signaling specificity|
|WO2008118324A2||Mar 20, 2008||Oct 2, 2008||Macrogenics, Inc.||Composition and method of treating cancer with an anti-uroplakin ib antibody|
|WO2008153926A2||Jun 5, 2008||Dec 18, 2008||Yale University||Inhibitors of receptor tyrosine kinases and methods of use thereof|
|WO2009091983A1 *||Jan 16, 2009||Jul 23, 2009||3M Innovative Properties Company||Methods for detecting an analyte|
|WO2009123894A2||Mar 25, 2009||Oct 8, 2009||Macrogenics, Inc.||Her2/neu-specific antibodies and methods of using same|
|WO2009151693A2||Mar 13, 2009||Dec 17, 2009||Eastern Virginia Medical School||Imaging mass spectrometry for improved prostate cancer diagnostics|
|WO2009151717A2||Mar 25, 2009||Dec 17, 2009||Macrogenics, Inc.||Bcr-complex-specific antibodies and methods of using same|
|WO2010021415A1||Aug 18, 2008||Feb 25, 2010||Seoul National University Industry Foundation||Method for controlling cancer metastasis or cancer cell migration by modulating the cellular level of lysyl trna synthetase|
|WO2010061393A1||Nov 30, 2009||Jun 3, 2010||Compugen Ltd.||He4 variant nucleotide and amino acid sequences, and methods of use thereof|
|WO2010065867A1||Dec 4, 2009||Jun 10, 2010||Pioneer Hi-Bred International, Inc.||Methods and compositions for enhanced yield by targeted expression of knotted1|
|WO2010067308A2||Dec 8, 2009||Jun 17, 2010||Compugen Ltd.|
|WO2010126670A2||Mar 26, 2010||Nov 4, 2010||Gojo Industries, Inc.||Compositions and methods for screening and using compounds antagonizing spore-surface interactions|
|WO2011012672A1||Jul 29, 2010||Feb 3, 2011||Pharnext||New diagnostic tools for alzheimer disease|
|WO2011067711A2||Nov 30, 2010||Jun 9, 2011||Compugen Ltd||Novel heparanase splice variant|
|WO2011106738A2||Feb 25, 2011||Sep 1, 2011||Fred Hutchinson Cancer Research Center||Use of tcr clonotypes as biomarkers for disease|
|WO2011109398A2||Mar 1, 2011||Sep 9, 2011||President And Fellows Of Harvard College||Methods and compositions for treatment of angelman syndrome and autism spectrum disorders|
|WO2012035518A1||Sep 16, 2011||Mar 22, 2012||Compugen Ltd.||Compositions and methods for treatment of drug resistant multiple myeloma|
|WO2012099576A1||Jan 18, 2011||Jul 26, 2012||Baxter International Inc.||Measurement of anti-amyloid antibodies in human blood|
|WO2012099897A1||Jan 17, 2012||Jul 26, 2012||Symbolics, Llc||Lateral flow assays using two dimensional features|
|WO2012139058A1||Apr 6, 2012||Oct 11, 2012||Biogen Idec Ma Inc.||BIOMARKERS PREDICTIVE OF THERAPEUTIC RESPONSIVENESS TO IFNβ AND USES THEREOF|
|WO2012160448A2||May 25, 2012||Nov 29, 2012||Innate Pharma, S.A.||Anti-kir antibodies for the treatment of inflammatory disorders|
|WO2013003112A1||Jun 19, 2012||Jan 3, 2013||The Jackson Laboratory||Methods and compositions for treatment of cancer and autoimmune disease|
|WO2013043012A2 *||Sep 24, 2012||Mar 28, 2013||Medicinal Bioconvergence Research Center||Novel use of leucyl trna synthetase|
|WO2013043012A3 *||Sep 24, 2012||May 16, 2013||Medicinal Bioconvergence Research Center||Novel use of leucyl trna synthetase|
|WO2014012077A1||Jul 12, 2013||Jan 16, 2014||Genisphere, Llc||Lateral flow assays using dna dendrimers|
|WO2014015076A1||Jul 17, 2013||Jan 23, 2014||Symbolics, Llc||Lateral flow assays using two dimensional features|
|WO2014071358A2||Nov 5, 2013||May 8, 2014||Foundation Medicine, Inc.||Novel ntrk1 fusion molecules and uses thereof|
|WO2014071406A1||Nov 5, 2013||May 8, 2014||Pronai Therapeutics, Inc.||Methods of using biomarkers for the treatment of cancer by modulation of bcl2|expression|
|WO2014081925A1||Nov 21, 2013||May 30, 2014||Agios Pharmaceuticals, Inc.||Glutamase inhibitors and method of use|
|WO2014113729A2||Jan 17, 2014||Jul 24, 2014||Foundation Mecicine, Inc.||Methods of treating cholangiocarcinoma|
|WO2015038978A1||Sep 12, 2014||Mar 19, 2015||Symbolics, Llc||Lateral flow assays using two dimensional test and control signal readout patterns|
|WO2015063604A2||Oct 31, 2014||May 7, 2015||Imberti Luisa||BIOMARKERS PREDICTIVE OF THERAPEUTIC RESPONSIVENESS TO IFNβ AND USES THEREOF|
|WO2015172083A1||May 8, 2015||Nov 12, 2015||Biogen Ma Inc.||Dimethylfumarate and prodrugs for treatment of multiple sclerosis|
|WO2015175424A1||May 11, 2015||Nov 19, 2015||Biogen Ma Inc.||Biomarkers predictive of lupus progression and uses thereof|
|WO2016026978A1||Aug 24, 2015||Feb 25, 2016||Universite Nice Sophia Antipolis||Methods and pharmaceutical compositions for treating drug addiction|
|WO2016057705A1||Oct 7, 2015||Apr 14, 2016||Novartis Ag||Biomarkers predictive of therapeutic responsiveness to chimeric antigen receptor therapy and uses thereof|
|U.S. Classification||435/5, 436/514, 436/527, 435/7.94, 436/538, 436/541, 435/7.93, 435/188, 436/530, 435/970|
|Cooperative Classification||Y10S435/97, G01N33/54386|
|Mar 23, 1981||AS||Assignment|
Owner name: AMERICAN HOSPITAL SUPPLY CORPORATION,ONE AMERICAN
Free format text: ASSIGNMENT OF ASSIGNORS INTEREST.;ASSIGNORS:GIEGEL JOSEPH L.;BROTHERTON MARY M.;REEL/FRAME:003841/0195
Effective date: 19810306
|Mar 2, 1987||AS||Assignment|
Owner name: BAXTER TRAVENOL LABORATORIES, INC. A CORP. OF DE
Free format text: MERGER;ASSIGNOR:AMERICAN HOSPITAL SUPPLY CORPORATION INTO;REEL/FRAME:004760/0345
Effective date: 19870126
|Oct 31, 1988||FPAY||Fee payment|
Year of fee payment: 4
|Jan 30, 1990||AS||Assignment|
Owner name: BAXTER INTERNATIONAL INC.
Free format text: CHANGE OF NAME;ASSIGNOR:BAXTER TRAVENOL LABORATORIES, INC., A CORP. OF DE;REEL/FRAME:005050/0870
Effective date: 19880518
|Mar 11, 1991||AS||Assignment|
Owner name: BAXTER DIAGNOSTICS INC.
Free format text: ASSIGNMENT OF ASSIGNORS INTEREST.;ASSIGNOR:BAXTER HEALTHCARE CORPORATION;REEL/FRAME:005712/0396
Effective date: 19900831
Owner name: BAXTER HEALTHCARE CORPORATION, A CORP. OF DE
Free format text: ASSIGNMENT OF ASSIGNORS INTEREST.;ASSIGNOR:BAXTER INTERNATIONAL INC., A CORP. OF DE;REEL/FRAME:005622/0729
Effective date: 19900831
|Sep 28, 1992||FPAY||Fee payment|
Year of fee payment: 8
|Jan 13, 1995||AS||Assignment|
Owner name: BANKERS TRUST COMPANY, NEW YORK
Free format text: ASSIGNMENT OF SECURITY INTEREST;ASSIGNORS:DIAGNOSTICS HOLDING, INC.;DADE INTERNATIONAL INC.;BARTELS, INC.;AND OTHERS;REEL/FRAME:007297/0204
Effective date: 19941220
|Jan 20, 1995||AS||Assignment|
Owner name: DADE INTERNATIONAL INC., ILLINOIS
Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:BAXTER DIAGNOSTICS INC.;REEL/FRAME:007272/0149
Effective date: 19941219
|Sep 30, 1996||FPAY||Fee payment|
Year of fee payment: 12
|Jun 1, 1998||AS||Assignment|
Owner name: DADE BEHRING INC., ILLINOIS
Free format text: CHANGE OF NAME;ASSIGNOR:DADE INTERNATIONAL INC.;REEL/FRAME:009328/0921
Effective date: 19980101
Owner name: BADE BEHRING INC., ILLINOIS
Free format text: CHANGE OF NAME;ASSIGNOR:DADE INTERNATIONAL INC.;REEL/FRAME:009297/0425
Effective date: 19980101
|Jun 8, 1998||AS||Assignment|
Owner name: DADE BEHRING INC., ILLINOIS
Free format text: CHANGE OF NAME;ASSIGNOR:DADE INTERNATIONAL INC.;REEL/FRAME:009267/0071
Effective date: 19980101
|Aug 20, 1998||AS||Assignment|
Owner name: DADE BEHRING INC., ILLINOIS
Free format text: CHANGE OF NAME;ASSIGNOR:DADE INTERNATIONAL INC.;REEL/FRAME:009405/0428
Effective date: 19980101
|Sep 15, 1999||AS||Assignment|
Owner name: BANKERS TRUST COMPANY, NEW YORK
Free format text: SECURITY AGREEMENT;ASSIGNOR:DADE BEHRING INC.;REEL/FRAME:010231/0085
Effective date: 19990629
|Nov 29, 2002||AS||Assignment|
Owner name: DEUTSCHE BANK AG, NEW YORK
Free format text: SECURITY INTEREST;ASSIGNOR:DADE BEHRING INC.;REEL/FRAME:013484/0739
Effective date: 20021003
|Mar 18, 2003||AS||Assignment|
Owner name: CHIMERA RESEARCH AND CHEMICAL INC., ILLINOIS
Free format text: PATENT RELEASE;ASSIGNOR:DEUTSCHE BANK TRUST COMPANY AMERICAS;REEL/FRAME:013821/0108
Effective date: 20021003
|May 4, 2005||AS||Assignment|
Owner name: DADE BEHRING INC., ILLINOIS
Free format text: RELEASE OF SECURITY INTEREST FOR PATENTS;ASSIGNOR:DEUTSCHE BANK AG, NEW YORK BRANCH;REEL/FRAME:015972/0363
Effective date: 20050426